...trend in funding and space in academic research labs has more students looking for other options. While graduate... skills for alternative careers. Science Careers has developed a web-based, career-planning platform called...your project may not advance beyond its initial stages. Help you build a network outside of the academic...

... a single step. Scorpion chemistry in particular has been reported to work well in the Lightcycler while...for all amplicons; if your amplicon is AT-rich or has a secondary structure, you may see a double peak ...being a common choice - the frequently used GAPDH has noted issues as a reference gene (Higgins, 2000; ...assumptions that match your data set, that is.) Yuan, 2006 has a good overview of different statistical analysis...reaction or on a rapid cycle machine. Melt Curves An advantage of Sybr chemistry is the ability to run a melt...prevent your reaction from reaching the plateau stage after exponential amplification, making it difficult...

...extraordinary collection of talent, of human knowledge, that has ever been gathered at the White House - with the ... not an excuse. Every psych test I’ve ever taken has declared me an introvert, and I identify as one.  ... on a leaderless team. Sometimes the leader just has to be the person to say “OK, guys, we need to have...anything out the discussion themselves, then one has to question whether that person should be on the ...the first place? Let’s think about some of the advantages of team-based work: Nobody knows everything anymore...

... Lab technician / Research assistant Typically has a Master's or Bachelor's degree Generally a short-term...talking to people, though, try to be conscious of who has your best interests at heart. A great piece of advice...vulnerable about my struggles and what I don’t know yet has helped me cultivate stronger relationships with my...academia or industry, so feel free to explore! Addgene has an entire page dedicated to different career options...position taken to transition to the next career stage (ex: grad school, med school, industry) Other...

...your data to Zooniverse. The Natural History Museum has downloadable resources to enable you to set up your... doors Getting involved in science communication has been the most rewarding step I have taken in my career...: Sign up for a Twitter account. Check out the hashtag #scicomm and start following science communication...local pub at a Pint of Science event. Or get on stage for a Science Slam – a sort of competitive TED talk...

...to host proteins. After the antibody of interest has been cloned into an expression plasmid, the plasmid...through open plasmid sharing, the CRISPR community has created hundreds of useful variants of Cas9. By providing...Antibodies Recombinant antibodies offer numerous advantages over polyclonal antibodies and traditional hybridomas...AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS...

...invention in the late 1990s, Gateway cloning technology has become very popular as a rapid and highly efficient...attL-containing entry vector.   Pro Tip: Addgene also has ready-made entry clones available for many popular...more about the Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway...and verify that your construct is functional. Advantages of the Gateway cloning method Compatibility and...JY, Chin CF (2015) Gateway Cloning Technology: Advantages and Drawbacks. Clon Transgen 4:138. doi:10.4172...

...requiring a double-stranded break. The Xiao Wang Lab has recently deposited a set of plasmids that helps identify...in human pluripotent stem cells, a cell type that has been difficult to modify in the past.   Figure...Cas13Y, two compact Cas13 proteins found in the metagenome datasets from hypersaline samples, were engineered...

...depositing lab. If your vector is large (>10 kb) or has a low copy number, try following protocols for low...easiest and clearest ways to determine if your sample has recombined. Choose a restriction enzyme that will...contains a mixture of the full plasmid and a smaller percentage of the recombined vector backbone.   Ready...

...TOPO-TA) cloning; however, the TOPO cloning technique has also be adapted for blunt end cloning. So how does...PCR products. Keep in mind that Taq polymerase has an error rate of about 1 in 3,500 bases. Typically...Resources on the Addgene Blog Perform Site Directed Mutagenesis by PCR Use REPLACR Mutatgenesis to Easily Generate...

...emotions, and even what therapeutic properties it has. As with other works of Dr. Sacks, Musicophilia includes... pretty biased as the founder, but our book club has become one of my favorite parts of Addgene’s culture...love a book series that features a young female protagonist, some clever magic, and a satisfying conclusion...

...effective than both AAV9 and AAV5, which has tropism for lung cells, and has increased overall transduction throughout...this definition: A systemic capsid is a capsid that has been engineered to have enhanced function after intravenous...both). Therefore, a high-efficiency systemic capsid has better efficacy than AAV9 and/or its intermediate...used in LY6A-nonpermissive mouse strains. **AAV1-X1 has not been tested in rats. For more information on ...neurons in the CNS (Chan et al., 2017). AAV-PHP.eB has frequently been used for screening candidate regulatory...depositing manuscript) across the brain. AAV-BI-28 has been tested in C57BL/6J mice and effectively transduces.... 4c, Jang et al., 2023; Brown et al., 2021.) It has a similar transduction rate to AAV-PHP.eB in the ...

... technique developed in David Liu’s lab in 2019, has added considerably more precision and flexibility... the canonical base editing window, base editing has higher efficiency and fewer indels than prime editing... repair (HDR) stimulated by double strand breaks has been widely used to generate precise changes. However...nicking of the unedited strand only after editing has occurred and reduces indels by 13-fold (Anzalone ... al., 2024; Yarnall et al., 2023) Prime editing has a lot of promise for treating genetic diseases. In...ones in Addgene’s repository later in the post. Advantages of prime editing Less constrained by PAM sequence...

...infect dividing and non-dividing cells.  Addgene has an extensive collection of lentiviral plasmids created... immortalized with adenoviral protein E1A, which has been shown to repress expression of HIV-1 LTRs (more...the deletion reduces the risk of insertional mutagenesis by limiting the interaction between viral LTR...potential for oncogenesis through insertional mutagenesis. These risks are largely based upon the vector...

...increase the sensitivity of detection (14-20). LDR has also been extended to multiplexed probe sets that...high-sensitivity multiplexed format (21, 27). This methodology has been used to rapidly screen buffer conditions for... 200 mM KCl at pH 8.5. This optimization method has been used internally at NEB to screen additional ...additional ligases, conditions, and formulations and has led to the development of the new HiFi Taq DNA Ligase....Numerous applications have been developed to take advantage of the high fidelity of Taq and other thermostable...

...CRISPR/Cas editing toolbox for C. elegans Recent work has further improved the efficiency of genome editing... (Farboud et al., 2019). Work from the Mello lab has dramatically improved RNP editing with linear repair...elegans (Ebbing et al., 2017). Because C. elegans has an AT-rich genome, it can be difficult to find an...whereas import of CRISPR technology into C. elegans has been a transformative innovation for our community...-CRISPR and co-conversion approaches have the advantage of being used in any genetic background, but require...

...the post) has a number of helpful suggestions.  Gels Gel issues usually come down to voltage and time....ladder. Unstained ladders contain either ‌a protein tag or an IgG binding site, allowing you to visualize...bands, your voltage is likely too high. If you’re seeing a smiley face at an appropriate voltage (usually...can happen with large proteins, try lowering the voltage and using a longer run time.   Samples can run ...packs can help keep the system cool, as can lower voltages.  If your small protein is over-transferring, ...transfer Increase transfer time Increase voltage Increase SDS/decrease alcohol in transfer buffer...Psi membrane Semi-dry transfer Decrease voltage Decrease transfer time Decrease SDS/increase...

... Addgene’s people, culture, and workplace. Rumor has it that a few Addgenies* may even be open to dancing...Social Media Are you following us on Twitter or Instagram (or Facebook or LinkedIn) but want to hear and...love sunshine and warm weather - check out our Instagram all summer long to see what kind of shenanigans...

...bind DNA targets without changing them. Thus, it has been widely adapted for a wide variety of applications...inhibition, among others. Over the past few years, dCas9 has also become a robust visualization tool to study ...guide sgRNA-Sirius-8XMS2 which will recruit the HaloTag with red fluorescent ligands to the loci of interest...

...supernatant during the production phase. However, this has not been well characterized as it is not a common...microvesicles as a vehicle for drug delivery that has recently gained interest special contexts, such as...researchers discovered that these particles have some advantages over the conventionally purified AAV vectors....